© New DMD (CA)n -repeats
(modified last January 25, 2001)
Contents
Per marker:
1. Marker details
2. Primer details
3. Allele frequencies
4. PCR-conditions
a. PCR ingredients
b. PCR-cycling
Markers:
DMDSTR07A has been contributed to these pages by Sander Kneppers, Dept. Human Genetics,
Leiden University Medical Center (E-mail sander@ruly46.MedFac.LeidenUniv.nl).
1. Marker details
Localization Xp21.1
Heterozygosity frequency 0.68
Length most frequent allele 226 bp
Length variation alleles 218-235 bp
Number of alleles found 12
2. Primer details
Name
Localization
Sequence
Forward primer
DMDSTR07A-F
intron 7: 22,068-22,085
ttctggttttctggtctg
Reverse primer
DMDSTR07A-R
intron 7: 22,298-22,279
gaatcaatctctctgtcaag
3. Allele frequencies
Allele number
Length in bp
Frequency
218
0.04
224
0.08
225
0.06
226
0.44
227
0.02
228
0.08
229
0.06
230
0.08
231
0.06
233
0.02
234
0.02
235
0.04
NOTE: frequencies determined on 50 chromosomes.
DMDSTR07B has been contributed to these pages by Sander Kneppers, Dept. Human Genetics,
Leiden University Medical Center (E-mail sander@ruly46.MedFac.LeidenUniv.nl).
1. Marker details
Localization Xp21.1
Heterozygosity frequency 0.44
Length most frequent allele 231 bp
Length variation alleles 227-241 bp
Number of alleles found 7
2. Primer details
Name
Localization
Sequence
Forward primer
DMDSTR07B-F
intron 7: 23,231-23,250
agctattatctgagaaagtc
Reverse primer
DMDSTR07B-R
intron 7: 23,462-23,443
tgaggtgaatttattaaggg
3. Allele frequencies
Allele number
Length in bp
Frequency
227
0.02
229
0.04
231
0.70
233
0.12
235
0.08
237
0.02
239
-
241
0.02
NOTE: frequencies determined on 50 chromosomes.
DMD-P20 DXS269) has been contributed to these pages by Sander Kneppers, Dept. Human
Genetics, Leiden University Medical Center (E-mail sander@ruly46.MedFac.LeidenUniv.nl).
1. Marker details
Localization Xp21.1
Heterozygosity frequency 0.59
Length most frequent allele 220 bp
Length variation alleles 216-241 bp
Number of alleles found 7
2. Primer details
Name
Localization
Sequence
Forward primer
DMD-P20F
intron 44: 2,948-2,967 (M86524)
acagaaggggaggtacagca
Reverse primer
DMD-P20R
intron 44: 3,185-3,164 (M86524)
gccacaacatatgcttgaaaga
3. Allele frequencies
Allele number
Length in bp
Frequency
216
0.09
220
0.59
224
0.04
228
0.17
230
0.04
239
0.06
241
0.02
NOTE: frequencies determined on 54 chromosomes.
a. PCR ingredients
Component
PCR-reaction
name concentration stock final concentration
amount in ”l
autoclaved water - -
15.4
SuperTaq buffer 10 * buffer 1 * buffer
2.5
dNTP's 2 mM 0.2 mM
2.5
primer mix 25 ”M of each primer 0.5 ”M
0.5
MgCl2 25 mM -
-
DMSO 100 % -
-
BSA 10 mg/ml -
-
Taq-polymerase 5U/”l 0.5 U
0.1
alpha-32 P-dCTP 1”Ci/”l -
-
DNA template 50 ng/”l 200 ng
4
final volume PCR 25
b. PCR-cycling
PCR step
temperature in șC
time in sec.
number of cycli
denaturation template 95
300
1
denaturation 94
30
30
annealing 55
30
elongation 72
30
final elongation 72
300
1
store 4
1
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DMD-P20 (DXS269)
(from Leiden)